Mechanism of tethered agonist-mediated signaling by polycystin-1.
Mechanism of tethered agonist-mediated signaling by polycystin-1.
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DOI：
10.1073/pnas.2113786119
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发表时间：
2022-05-10
影响因子：
11.1
通讯作者：
中科院分区：
文献类型：
--
作者：
多囊素-1 （PC1）突变是常染色体显性多囊肾病（ADPKD）的主要原因（85%的病例），这是肾衰竭的第四大原因。PC1被认为是一种非典型G蛋白偶联受体，但PC1调节G蛋白信号传导的机制仍然知之甚少。PC1的ADPKD突变中有很大一部分编码了一种成熟缺陷或功能降低的蛋白质，这种蛋白质可能适合于功能修复。在这项工作中，我们结合了互补的生化和细胞分析实验以及加速的分子模拟，揭示了PC1 c端片段激活的变构转导途径。我们的发现将有助于未来针对PC1信号功能的合理药物设计。多囊蛋白-1 （PC1）是一种重要的异常G蛋白偶联受体（GPCR），具有11个跨膜结构域，其突变占常染色体显性多囊肾病（ADPKD）病例的85%。PC1与粘附gpcr具有多种特性。其中包括一个GPCR蛋白水解位点，它可以自动催化地将这些蛋白质分裂成细胞外、n端和膜嵌入的c端片段（CTF），以及CTF n端柄内的拴系激动剂（TA），它被认为可以激活信号传导。然而，TA激活PC1的机制尚不清楚。在这里，我们将编码野生型、无茎型和三种不同ADPKD茎变异体的PC1 CTF表达构建体的功能细胞信号传导实验与全原子高斯加速分子动力学（GaMD）模拟相结合，研究ta介导的信号激活。GaMD模拟计算的残基运动和自由能分布的相关性与PC1 CTF野生型和茎秆变体的差异信号传导能力相关。他们提出了一种变构机制，涉及连接茎的残基相互作用，多囊毒素（TOP）结构域的四方开放，以及ta介导的PC1 CTF激活的假定孔环。通过诱变实验验证了gmd模拟预测的N3074-S3585和R3848-E4078等关键相互作用残基。总之，这些互补分析提供了对ta介导的PC1 CTF信号激活机制的见解，这将对未来针对PC1的合理药物设计具有重要意义。
Mutations of polycystin-1 (PC1) are the major cause (85% of cases) of autosomal dominant polycystic kidney disease (ADPKD), which is the fourth leading cause of kidney failure. PC1 is thought to function as an atypical G protein-coupled receptor, yet the mechanism by which PC1 regulates G-protein signaling remains poorly understood. A significant portion of ADPKD mutations of PC1 encode a protein with defects in maturation or reduced function that may be amenable to functional rescue. In this work, we have combined complementary biochemical and cellular assay experiments and accelerated molecular simulations, which revealed an allosteric transduction pathway in activation of the PC1 C-terminal fragment. Our findings will facilitate future rational drug design efforts targeting the PC1 signaling function. Polycystin-1 (PC1) is an important unusual G protein-coupled receptor (GPCR) with 11 transmembrane domains, and its mutations account for 85% of cases of autosomal dominant polycystic kidney disease (ADPKD). PC1 shares multiple characteristics with Adhesion GPCRs. These include a GPCR proteolysis site that autocatalytically divides these proteins into extracellular, N-terminal, and membrane-embedded, C-terminal fragments (CTF), and a tethered agonist (TA) within the N-terminal stalk of the CTF that is suggested to activate signaling. However, the mechanism by which a TA can activate PC1 is not known. Here, we have combined functional cellular signaling experiments of PC1 CTF expression constructs encoding wild type, stalkless, and three different ADPKD stalk variants with all-atom Gaussian accelerated molecular dynamics (GaMD) simulations to investigate TA-mediated signaling activation. Correlations of residue motions and free-energy profiles calculated from the GaMD simulations correlated with the differential signaling abilities of wild type and stalk variants of PC1 CTF. They suggested an allosteric mechanism involving residue interactions connecting the stalk, Tetragonal Opening for Polycystins (TOP) domain, and putative pore loop in TA-mediated activation of PC1 CTF. Key interacting residues such as N3074–S3585 and R3848–E4078 predicted from the GaMD simulations were validated by mutagenesis experiments. Together, these complementary analyses have provided insights into a TA-mediated activation mechanism of PC1 CTF signaling, which will be important for future rational drug design targeting PC1.
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影响因子：
11.4
作者：
Arac, Demet;Boucard, Antony A.;Bolliger, Marc F.;Nguyen, Jenna;Soltis, S. Michael;Suedhof, Thomas C.;Brunger, Axel T.
通讯作者：
Brunger, Axel T.
影响因子：
5.5
作者：
Miao Y;Feher VA;McCammon JA
通讯作者：
McCammon JA
影响因子：
4.8
作者：
Kishore, Ayush;Hall, Randy A.
通讯作者：
Hall, Randy A.
影响因子：
5.8
作者：
Nazarko O;Kibrom A;Winkler J;Leon K;Stoveken H;Salzman G;Merdas K;Lu Y;Narkhede P;Tall G;Prömel S;Araç D
通讯作者：
Araç D
DOI：
10.1016/j.bbrc.2015.07.020
发表时间：
2015-08-28
影响因子：
3.1
作者：
Demberg, Lilian M.;Rothemund, Sven;Liebscher, Ines
通讯作者：
Liebscher, Ines